Prospective uses of recombinant Lactococcus lactis expressing both listeriolysin O and mutated internalin A from Listeria monocytogenes as a tool for DNA vaccination

In the present report, we describe how Lactococcus lactis strains ectopically express the internalin-A protein from Listeria monocytogenes at the cell surface, situated specifically under the bacterial septum region. Such findings provide relevant insights into the spatial expression pattern of exogenous genes in lactic bacteria.

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Expression of two Listeria monocytogenes antigens (P60 and LLO) in Lactococcus lactis and examination for use as live vaccine vectors

Journal of Medical Microbiology ◽

10.1099/jmm.0.018770-0 ◽

2010 ◽

Vol 59 (8) ◽

pp. 904-912 ◽

Cited By ~ 20

Author(s):

Mohammed Bahey-El-Din ◽

Pat G. Casey ◽

Brendan T. Griffin ◽

Cormac G. M. Gahan

Keyword(s):

Immune Response ◽

Listeria Monocytogenes ◽

Lactococcus Lactis ◽

Protective Efficacy ◽

Oral Immunization ◽

Listeriolysin O ◽

Oral Vaccination ◽

Vaccine Vectors ◽

Food Borne ◽

Presentation Pathway

Listeria monocytogenes is a food-borne intracellular pathogen that mainly infects pregnant and immunocompromised individuals. The pore-forming haemolysin listeriolysin O (LLO), the main virulence factor of Listeria monocytogenes, allows bacteria to escape from the harsh environment of the phagosome to the cytoplasm of the infected cell. This leads to processing of bacterial antigens predominantly through the cytosolic MHC class I presentation pathway. We previously engineered the food-grade bacterium Lactococcus lactis to express LLO and demonstrated an LLO-specific CD8+ response upon immunization of mice with the engineered L. lactis vaccine strains. In the present work, we examined the immune response and protective efficacy of an L. lactis strain co-expressing LLO and a truncated form of the listerial P60 antigen (tP60). Oral immunization revealed no significant protection against listeriosis with L. lactis expressing LLO, tP60 or the combined LLO/tP60. In contrast, intraperitoneal vaccination induced an LLO-specific CD8+ immune response with LLO-expressing L. lactis but no significant improvement in protection was observed following vaccination with the combined LLO/tP60 expressing L. lactis strain. This may be due to the low level of tP60 expression in the LLO/tP60 strain. These results demonstrate the necessity for improved oral vaccination strategies using LLO-expressing L. lactis vaccine vectors.

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Virulence potential of Listeria monocytogenes strains recovered from pigs in Spain

Veterinary Record ◽

10.1136/vr.105945 ◽

2020 ◽

Vol 187 (11) ◽

pp. e101-e101

Author(s):

Jaime Gómez-Laguna ◽

Fernando Cardoso-Toset ◽

Jazmín Meza-Torres ◽

Javier Pizarro-Cerdá ◽

Juan J Quereda

Keyword(s):

Listeria Monocytogenes ◽

Virulence Factors ◽

Bacterial Pathogen ◽

Listeriolysin O ◽

Actin Assembly ◽

Virulence Potential ◽

Serotype 1 ◽

Internalin B ◽

Meat Samples ◽

Internalin A

BackgroundListeria monocytogenes is a foodborne bacterial pathogen that causes listeriosis, an infectious disease in animals and people, with pigs acting as asymptomatic reservoirs. In August 2019 an outbreak associated with the consumption of pork meat caused 222 human cases of listeriosis in Spain. Determining the diversity as well as the virulence potential of strains from pigs is important to public health.MethodsThe behaviour of 23 L monocytogenes strains recovered from pig tonsils, meat and skin was compared by studying (1) internalin A, internalin B, listeriolysin O, actin assembly-inducing protein and PrfA expression levels, and (2) their invasion and intracellular growth in eukaryotic cells.ResultsMarked differences were found in the expression of the selected virulence factors and the invasion and intracellular replication phenotypes of L monocytogenes strains. Strains obtained from meat samples and belonging to serotype 1/2a did not have internalin A anchored to the peptidoglycan. Some strains expressed higher levels of the studied virulence factors and invaded and replicated intracellularly more efficiently than an epidemic L monocytogenes reference strain (F2365).ConclusionThis study demonstrates the presence of virulent L monocytogenes strains with virulent potential in pigs, with valuable implications in veterinary medicine and food safety.

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In vitro and in vivo characterization of DNA delivery using recombinant Lactococcus lactis expressing a mutated form of L. monocytogenes Internalin A

BMC Microbiology ◽

10.1186/1471-2180-12-299 ◽

2012 ◽

Vol 12 (1) ◽

pp. 299 ◽

Cited By ~ 24

Author(s):

Marcela de Azevedo ◽

Jurgen Karczewski ◽

François Lefévre ◽

Vasco Azevedo ◽

Anderson Miyoshi ◽

...

Keyword(s):

Lactococcus Lactis ◽

Dna Delivery ◽

Recombinant Lactococcus Lactis ◽

In Vivo Characterization ◽

Internalin A

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Lactococcus lactis and Lactobacillus salivarius differently modulate early immunological response of Wistar rats co-administered with Listeria monocytogenes

Beneficial Microbes ◽

10.3920/bm2017.0007 ◽

2017 ◽

Vol 8 (5) ◽

pp. 809-822 ◽

Cited By ~ 7

Author(s):

J. Lukic ◽

I. Jancic ◽

N. Mirkovic ◽

B. Bufan ◽

J. Djokic ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Mrna Expression ◽

Lactococcus Lactis ◽

Wistar Rats ◽

Immunological Response ◽

Listeriolysin O ◽

Mesenteric Lymph Nodes ◽

Lactobacillus Salivarius ◽

Cd69 Expression ◽

Immunological Effects

In the light of the increasing resistance of bacterial pathogens to antibiotics, one of the main global strategies in applied science is development of alternative treatments, which would be safe both for the host and from the environmental perspective. Accordingly, the aim of this study was to test whether two lactic acid bacteria (LAB) strains, Lactococcus lactis BGBU1-4 and Lactobacillus salivarius BGHO1, could be applied as safe supplements for Listeria infection. Two major research objectives were set: to compare the effects of BGBU1-4 and BGHO1 on early immune response in gut tissue of Wistar rats co-administered with Listeria monocytogenes ATCC19111 and next, to test how this applies to their usage as therapeutics in acute ATCC19111 infection. Intestinal villi (IV), Peyer’s patches (PP) and mesenteric lymph nodes (MLN) were used for the analysis. The results showed that BGHO1 increased the mRNA expression of innate immune markers CD14, interleukin (IL)-1β and tumour necrosis factor (TNF)-α in PP and IV, and, in parallel, caused a decrease of listeriolysin O (LLO) mRNA expression in same tissues. In MLN of BGHO1 treated rats, LLO expression was increased, along with an increase of the expression of OX-62 mRNA and CD69, pointing to the activation of adaptive immunity. On the other hand, in BGBU1-4 treated rats, there was no reduction of LLO mRNA expression and no induction of innate immunity markers in intestinal tissue. Additionally, CD14 and IL-1β, as well as LLO, but not OX-62 mRNA and CD69 expression, were elevated in MLN of BGBU1-4 treated rats. However, when applied therapeutically, both, BGBU1-4 and BGHO1, lowered Listeria count in spleens of infected rats. Our results not only reveal the potential of LAB to ameliorate Listeria infections, but suggest different immunological effects of two different LAB strains, both of which could be effective in Listeria elimination.

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Lactococcus lactis-expressing listeriolysin O (LLO) provides protection and specific CD8+ T cells against Listeria monocytogenes in the murine infection model

Vaccine ◽

10.1016/j.vaccine.2008.07.047 ◽

2008 ◽

Vol 26 (41) ◽

pp. 5304-5314 ◽

Cited By ~ 43

Author(s):

Mohammed Bahey-El-Din ◽

Pat G. Casey ◽

Brendan T. Griffin ◽

Cormac G.M. Gahan

Keyword(s):

T Cells ◽

Listeria Monocytogenes ◽

Lactococcus Lactis ◽

Cd8 T Cells ◽

Infection Model ◽

Listeriolysin O

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Lactiplantibacillus plantarum subsp. plantarum and Fructooligosaccharides Combination Inhibits the Growth, Adhesion, Invasion, and Virulence of Listeria monocytogenes

Foods ◽

10.3390/foods11020170 ◽

2022 ◽

Vol 11 (2) ◽

pp. 170

Author(s):

Qingli Dong ◽

Xinxin Lu ◽

Binru Gao ◽

Yangtai Liu ◽

Muhammad Zohaib Aslam ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Incubation Temperature ◽

Foodborne Pathogen ◽

Untreated Group ◽

Listeriolysin O ◽

Regulatory Factor ◽

Gene Expressions ◽

Internalin B ◽

Serotype 4B ◽

Internalin A

Listeria monocytogenes is a foodborne pathogen responsible for many food outbreaks worldwide. This study aimed to investigate the single and combined effect of fructooligosaccharides (FOS) and Lactiplantibacillus plantarum subsp. plantarum CICC 6257 (L. plantarum) on the growth, adhesion, invasion, and virulence of gene expressions of Listeria monocytogenes 19112 serotype 4b (L. monocytogenes). Results showed that L. plantarum combined with 2% and 4% (w/v) FOS significantly (p < 0.05) inhibited the growth of L. monocytogenes (3–3.5 log10 CFU/mL reduction) at the incubation temperature of 10 °C and 25 °C. Under the same combination condition, the invasion rates of L. monocytogenes to Caco-2 and BeWo cells were reduced more than 90% compared to the result of the untreated group. After L. plantarum was combined with the 2% and 4% (w/v) FOS treatment, the gene expression of actin-based motility, sigma factor, internalin A, internalin B, positive regulatory factor A, and listeriolysin O significantly (p < 0.05) were reduced over 91%, 77%, 92%, 89%, 79%, and 79% compared to the result of the untreated group, respectively. The inhibition level of the L. plantarum and FOS combination against L. monocytogenes was higher than that of FOS or L. plantarum alone. Overall, these results indicated that the L. plantarum and FOS combination might be an effective formula against L. monocytogenes.

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